Apparatus for screening compound libraries

Abstract

Disclosed are apparatus for screening compound libraries using frontal chromatography in combination with mass spectrometry to identify and rank those members of the library that bind to a target receptor. The apparatus of this invention also permit a compound library to be rapidly screened to determine if any member of the library has a higher affinity for the target receptor relative to a pre-selected indicator compound.

What is claimed is:  
     
       1. An apparatus for screening a plurality of compound libraries to determine the relative or absolute affinity of a plurality of putative ligands in each library to a target receptor, which apparatus comprises: 
       (a) a plurality of columns each comprising a target receptor optionally attached to a solid phase support and having a inflow end and an outflow end, wherein each of said columns is capable of independently having a compound library comprising a plurality of putative ligands continuously applied thereto under frontal chromatography conditions whereby the target receptor is continuously contacted with the compound library to produce an effluent from the outflow end of the column;  
       (b) a plurality of first reservoir connected to the inflow end of one of the columns for continuously applying the compound library to the columns;  
       (c) a mass spectrometer connected to the outflow end of each of said columns for intermittently analyzing the effluent from each of the column.  
     
     
       2. The apparatus of  claim 1 , wherein said apparatus further comprises: 
       (d) a plurality of second reservoirs each connected to the inflow end of one of the columns for applying either (i) a mixture comprising the compound library and one or more indicator compounds, (ii) one or more indicator compounds, or (iii) a buffer solution to the column.  
     
     
       3. The apparatus of  claim 1 , wherein said apparatus further comprises: 
       (e) a third reservoir connected to the outflow end of each of the columns for supplying a supplemental diluent to the effluent from each column before analysis by the mass spectrometer.  
     
     
       4. The apparatus of  claim 1 , wherein said apparatus comprises from 2 to about 100 columns. 
     
     
       5. The apparatus of  claim 4 , wherein said apparatus comprises from 3 to about 50 columns. 
     
     
       6. The apparatus of  claim 5 , wherein said apparatus comprises from 5 to about 10 columns. 
     
     
       7. The apparatus of  claim 1 , wherein each column is intermittently monitored for a period of about 0.5 seconds to about 10 seconds before switching to the next column. 
     
     
       8. The apparatus of  claim 7 , wherein each column is intermittently monitored for about 1 second to about 5 seconds before switching to the next column. 
     
     
       9. The apparatus of  claim 1 , wherein the column has an internal diameter ranging from about 10 μm to about 4.6 μmm. 
     
     
       10. The apparatus of  claim 9 , wherein the column has an internal diameter of from about 100 μm to about 250 μm. 
     
     
       11. The apparatus of  claim 1 , wherein the column has a length of from about 1 cm to about 30 cm. 
     
     
       12. The apparatus of  claim 1 , wherein the column has a length of from about 2 cm to about 20 cm. 
     
     
       13. The apparatus of  claim 1 , wherein the target receptor is selected from the group consisting of proteins, glycoproteins, glycosaminoglycans, proteoglycans, integrins, enzymes, lectins, selecting, cell-adhesion molecules, toxins, bacterial pili, transport proteins, receptors involved in signal transduction or hormone-binding, hormones, antibodies, major histocompatability complexes, immunoglobulin superfamilies, cadherins, DNA or DNA fragments, RNA and RNA fragments, whole cells, tissues, bacteria, fungi, viruses, parasites, preons, and synthetic analogs or derivatives thereof. 
     
     
       14. The apparatus of  claim 1 , wherein the target receptor is bound to a solid phase support. 
     
     
       15. The apparatus of  claim 14 , wherein the target receptor is covalently bound to the solid phase support or bound via biotin-avidin or biotin-streptavidin binding. 
     
     
       16. The apparatus of  claim 14 , wherein the solid phase support is selected from the group consisting of resin beads, glass beads, silica chips, silica capillaries and agarose. 
     
     
       17. The apparatus of  claim 1 , wherein the column contains from about 1 pmol to about 10 nmol of target receptor active sites. 
     
     
       18. The apparatus of  claim 1 , wherein the mass spectrometer is an electrospray mass spectrometer.